Method of using cyclooxygenase-2 inhibitors in the treatment and prevention of neoplasia

ABSTRACT

This invention relates to the use of cyclooxygenase-2 inhibitors or derivatives thereof in preventing and treating neoplasia. In particular, the invention describes the method of preventing and treating epithelial cell neoplasia in a subject, said method comprising treating the subject with a therapeutically-effective amount of a compound of Formula I.wherein A, R2 and R3 are as described in the specification.

This is a continuation of application Ser. No. 09/390,459 filed Sep. 7,1999 now abandoned which is a continuation of application Ser. No.08/949,922 filed Oct. 14, 1997, now U.S. Pat. No. 5,772,986.

FIELD OF THE INVENTION

This invention is in the field of the prevention and treatment ofneoplasia. More specifically, this invention relates to the use ofcyclooxygenase-2 inhibitors or derivatives thereof in preventing andtreating neoplasia.

BACKGROUND OF THE INVENTION

Prostaglandins play a major role in the inflammation process and theinhibition of prostaglandin production, especially production of PGG₂,PGH₂ and PGE₂, has been a common target of anti-inflammatory drugdiscovery. However, common non-steroidal anti-inflammatory drugs(NSAID's) that are active in reducing the prostaglandin-induced pain andswelling associated with the inflammation process are also active inaffecting other prostaglandin-regulated processes not associated withthe inflammation process. Thus, use of high doses of most common NSAID'scan produce severe side effects, including life threatening ulcers, thatlimit their therapeutic potential. An alternative to NSAID's is the useof corticosteroids, which also produce adverse effects, especially whenlong term therapy is involved.

NSAIDs have been found to prevent the production of prostaglandins byinhibiting enzymes in the human arachidonic acid/prostaglandin pathway,including the enzyme cyclooxygenase (COX). The recent discovery of aninducible enzyme associated with inflammation (named “cyclooxygenase-2(COX-2)” or “prostaglandin G/H synthase II”) provides a viable target ofinhibition which more effectively reduces inflammation and producesfewer and less drastic side effects.

Compounds which selectively inhibit cyclooxygenase-2 have been describedin U.S. Pat. Nos. 5,380,738, 5,344,991, 5,393,790, 5,434,178, 5,474,995,5,510,368 and WO documents WO96/06840, WO96/03388, WO96/03387,WO96/25405, WO95/15316, WO94/15932, WO94/27980, WO95/00501, WO94/13635,WO94/20480, and WO94/26731.

Neoplastic disease states are serious and oftentimes life-threateningconditions. These neoplastic diseases, which are characterized byrapidly-proliferating cell growth, continue to be the subject ofworldwide research efforts directed toward the identification oftherapeutic agents which are effective in the treatment thereof.Effective therapeutic agents prolong the survivability of the patient,inhibit the rapidly-proliferating cell growth associated with theneoplasm, or effect a regression of the neoplasm. Research in this areais primarily focused toward identifying agents which would betherapeutically effective in humans and other mammals.

Recently, the presence of COX-2 has been observed in neoplastic disease.See Masanobu Oshima et al. (Cell, 87, 803-809 (1996); and MichelleParret et al. (International Journal of Oncology, 10, 503-507 (1997).

[Pyrazol-1-yl]benzenesulfonamides have been described as inhibitors ofcyclooxygenase-2 and have shown promise in the treatment ofinflammation, arthritis, and pain, with minimal side effects inpre-clinical and clinical trials. Their use for preventing colon cancerhas been described in U.S. Pat. No. 5,466,823. However, their use fortreating colon cancer or for treating or preventing other neoplasias hasnot been previously described.

The present invention is directed to the use of inhibitors ofcyclooxygenase-2 for the treatment and prevention of neoplasias.Conjunctive treatment of a selective cyclooxygenase-2 inhibitor withother neoplastic agents produces a synergistic effect or alternativelyreduces the toxic side effects associated with chemotherapy by reducingthe concentration of the side effect-causing agent needed fortherapeutic efficacy.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a method for treating or preventing aneoplasia that produces a prostaglandin in a subject in need of suchtreatment or prevention, the method comprises treating the subject witha therapeutically effective amount of a cyclooxygenase-2 inhibitor orderivative thereof.

The term “treatment” includes partial or total inhibition of theneoplasia growth, spreading or metastasis, as well as partial or totaldestruction of the neoplasia cells.

The term “prevention” includes either preventing the onset of clinicallyevident neoplasia altogether or preventing the onset of a preclinicallyevident stage of neoplasia in individuals at risk. Also intended to beencompassed by this definition is the prevention of initiation formalignant cells or to arrest or reverse the progression of premalignantcells to malignant cells. This includes prophylactic treatment of thoseat risk of developing the neoplasia.

The phrase “therapeutically-effective” is intended to qualify the amountof each agent which will achieve the goal of improvement in diseaseseverity and the frequency of incidence over treatment of each agent byitself, while avoiding adverse side effects typically associated withalternative therapies.

The term “subject” for purposes of treatment includes any human ormammal subject who has any one of the known neoplasias, and preferablyis a human subject. For methods of prevention, the subject is any humanor animal subject, and preferably is a human subject who is at risk forobtaining an epithelium cell-derived neoplasia. The subject may be atrisk due to exposure to carcinogenic agents, being geneticallypredisposed to have the neoplasia, and the like.

The term “neoplasia” includes neoplasia that produce prostaglandins orexpress a cyclooxygenase, including both benign and cancerous tumors,growths and polyps.

In the method above, the neoplasia that produce prostaglandins includebrain cancer, bone cancer, epithelial cell-derived neoplasia (epithelialcarcinoma) such as basal cell carcinoma, adenocarcinoma,gastrointestinal cancer such as lip cancer, mouth cancer, esophogealcancer, small bowel cancer and stomach cancer, colon cancer, livercancer, bladder cancer, pancreas cancer, ovary cancer, cervical cancer,lung cancer, breast cancer and skin cancer, such as squamus cell andbasal cell cancers, prostate cancer, renal cell carcinoma, and otherknown cancers that effect epithelial cells throughout the body.Preferably, neoplasia is selected from gastrointestinal cancer, livercancer, bladder cancer, pancreas cancer, ovary cancer, prostate cancer,cervical cancer, lung cancer, breast cancer and skin cancer, such assquamus cell and basal cell cancers. The COX-2 inhibitors can also beused to treat the fibrosis which occurs with radiation therapy. Themethod can be used to treat subjects having adenomatous polyps,including those with familial adenomatous polyposis (FAP). Additionally,the method can be used to prevent polyps from forming in patients atrisk of FAP.

Inhibitors of the cyclooxygenase pathway in the metabolism ofarachidonic acid used in the prevention and treatment of epithelial cellderived neoplasias may inhibit enzyme activity through a variety ofmechanisms. By the way of example, the inhibitors used in the methodsdescribed herein may block the enzyme activity directly by acting as asubstrate for the enzyme. The use of cyclooxygenasse-2 selectiveinhibitors is highly advantageous in that they minimize the gastric sideeffects that can occur with non-selective NSAID's, especially whereprolonged prophylactic treatment is expected.

The term “cyclooxygenase-2 inhibitor” denotes a compound able to inhibitcyclooxygenase-2 without significant inhibition of cyclooxygenase-1.Preferably, it includes compounds which have a cyclooxygenase-2 IC₅₀ ofless than about 0.2 μM, and also have a selectivity ratio ofcyclooxygenase-2 inhibition over cyclooxygenase-1 inhibition of at least50, and more preferably of at least 100. Even more preferably, thecompounds have a cyclooxygenase-1 IC₅₀ of greater than about 1 μM, andmore preferably of greater than 10 μM. Pyrazoles can be prepared bymethods described in WO95/15316, WO95/15315 and WO96/03385. Thiopheneanalogs can be prepared by methods described in WO95/00501 andWO94/15932. Oxazoles can be prepared by the methods described in PCTdocuments WO95/00501 and WO94/27980. Isoxazoles can be prepared by themethods described in WO96/25405. Imidazoles can be prepared by themethods described in WO96/03388 and WO96/03387. Cyclopentenecyclooxygenase-2 inhibitors can be prepared by the methods described inU.S. Pat. No. 5,344,991 and WO 95/00501. Terphenyl compounds can beprepared by the methods described in WO96/16934. Thiazole compounds canbe prepared by the methods described in WO96/03392. Pyridine compoundscan be prepared by the methods described in WO96/24584 and WO96/24585.

The method provided herein relates to the use of cyclooxygenase-2inhibitors or derivatives thereof in the prevention and treatment ofderived neoplasias. In the preferred embodiments, the cycclooxygenase-2compound is selected from the compounds of Formula I

wherein A is a substituent selected from partially unsaturated orunsaturated heterocyclyl and partially unsaturated or unsaturatedcarbocyclic rings;

wherein R¹ is at least one substituent selected from heterocyclyl,cycloalkyl, cycloalkenyl and aryl, wherein R¹ is optionally substitutedat a substitutable position with one or more radicals selected fromalkyl, haloalkyl, cyano, carboxyl, alkoxycarbonyl, hydroxyl,hydroxyalkyl, haloalkoxy, amino, alkylamino, arylamino, nitro,alkoxyalkyl, alkylsulfinyl, halo, alkoxy and alkylthio;

wherein R² is methyl or amino; and

wherein R³ is a radical selected from hydrido, halo, alkyl, alkenyl,alkynyl, oxo, cyano, carboxyl, cyanoalkyl, heterocyclyloxy, alkyloxy,alkylthio, alkylcarbonyl, cycloalkyl, aryl, haloalkyl, heterocyclyl,cycloalkenyl, aralkyl, heterocyclylalkyl, acyl, alkylthioalkyl,hydroxyalkyl, alkoxycarbonyl, arylcarbonyl, aralkylcarbonyl, aralkenyl,alkoxyalkyl, arylthioalkyl, aryloxyalkyl, aralkylthioalkyl,aralkoxyalkyl, alkoxyaralkoxyalkyl, alkoxycarbonylalkyl, aminocarbonyl,aminocarbonylalkyl, alkylaminocarbonyl, N-arylaminocarbonyl,N-alkyl-N-arylaminocarbonyl, alkylaminocarbonylalkyl, carboxyalkyl,alkylamino, N-arylamino, N-aralkylamino, N-alkyl-N-aralkylamino,N-alkyl-N-arylamino, aminoalkyl, alkylaminoalkyl, N-arylaminoalkyl,N-aralkylaminoalkyl, N-alkyl-N-aralkylaminoalkyl,N-alkyl-N-arylaminoalkyl, aryloxy, aralkoxy, arylthio, aralkylthio,alkylsulfinyl, alkylsulfonyl, aminosulfonyl, alkylaminosulfonyl,N-arylaminosulfonyl, arylsulfonyl, N-alkyl-N-arylaminosulfonyl; or apharmaceutically-acceptable salt thereof.

A preferred class of compounds which inhibit cyclooxygenase-2 consistsof compounds of Formula I wherein A is selected from 5- or 6-memberpartially unsaturated heterocyclyl, 5- or 6-member unsaturatedheterocyclyl, 9- or 10-member unsaturated condensed heterocyclyl, lowercycloalkenyl and phenyl; wherein R¹ is selected from 5- and 6-memberedheterocyclyl, lower cycloalkyl, lower cycloalkenyl and aryl selectedfrom phenyl, biphenyl and naphthyl, wherein R¹ is optionally substitutedat a substitutable position with one or more radicals selected fromlower alkyl, lower haloalkyl, cyano, carboxyl, lower alkoxycarbonyl,hydroxyl, lower hydroxyalkyl, lower haloalkoxy, amino, lower alkylamino,phenylamino, lower alkoxyalkyl, lower alkylsulfinyl, halo, lower alkoxyand lower alkylthio; wherein R² is methyl or amino; and wherein R³ is aradical selected from hydrido, oxo, cyano, carboxyl, loweralkoxycarbonyl, lower carboxyalkyl, lower cyanoalkyl, halo, lower alkyl,lower alkyloxy, lower cycloalkyl, phenyl, lower haloalkyl, 5- or6-membered heterocyclyl, lower hydroxylalkyl, lower aralkyl, acyl,phenylcarbonyl, lower alkoxyalkyl, 5- or 6-membered heteroaryloxy,aminocarbonyl, lower alkylaminocarbonyl, lower alkylamino, loweraminoalkyl, lower alkylaminoalkyl, phenyloxy, and lower aralkoxy; or apharmaceutically-acceptable salt thereof.

A more preferred class of compounds which inhibit cyclooxygenase-2consists of compounds of Formula I wherein A is selected from oxazolyl,isoxazolyl, furyl, thienyl, dihydrofuryl, pyrrolyl, pyrazolyl,thiazolyl, imidazolyl, isothiazolyl, benzofuryl, cyclopentenyl,cyclopentadienyl, phenyl, and pyridyl; wherein R¹ is selected frompyridyl optionally substituted at a substitutable position with one ormore methyl radicals, and phenyl optionally substituted at asubstitutable position with one or more radicals selected from methyl,ethyl, isopropyl, butyl, tert-butyl, isobutyl, pentyl, hexyl,fluoromethyl, difluoromethyl, trifluoromethyl, cyano, carboxyl,methoxycarbonyl, ethoxycarbonyl, hydroxyl, hydroxymethyl,trifluoromethoxy, amino, N-methylamino, N,N-dimethylamino, N-ethylamino,N,N-dipropylamino, N-butylamino, N-methyl-N-ethylamino, phenylamino,methoxymethyl, methylsulfinyl, fluoro, chloro, bromo, methoxy, ethoxy,propoxy, n-butoxy, pentoxy, and methylthio; wherein R² is methyl oramino; and wherein R³ is a radical selected from hydrido, oxo, cyano,carboxyl, methoxycarbonyl, ethoxycarbonyl, carboxypropyl, carboxymethyl,carboxyethyl, cyanomethyl, fluoro, chloro, bromo, methyl, ethyl,isopropyl, butyl, tert-butyl, isobutyl, pentyl, hexyl, difluoromethyl,trifluoromethyl, pentafluoroethyl, heptafluoropropyl, difluoroethyl,difluoropropyl, methoxy, ethoxy, propoxy, n-butoxy, pentoxy, cyclohexyl,phenyl, pyridyl, thienyl, thiazolyl, oxazolyl, furyl, pyrazinyl,hydroxylmethyl, hydroxylpropyl, benzyl, formyl, phenylcarbonyl,methoxymethyl, furylmethyloxy, aminocarbonyl, N-methylaminocarbonyl,N,N-dimethylaminocarbonyl, N,N-dimethylamino, N-ethylamino,N,N-dipropylamino, N-butylamino, N-methyl-N-ethylamino, aminomethyl,N,N-dimethylaminomethyl, N-methyl-N-ethylaminomethyl, benzyloxy, andphenyloxy; or a pharmaceutically-acceptable salt thereof.

A family of specific compounds of particular interest within Formula Iconsists of compounds and pharmaceutically-acceptable salts thereof asfollows:

5-(4-fluorophenyl)-1-[4-(methylsulfonyl)phenyl]-3-(trifluoromethyl)pyrazole;

4-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]-1-phenyl-3-(trifluoromethyl)pyrazole;

4-(5-(4-chlorophenyl)-3-(4-methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide

4-(3,5-bis(4-methylphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;

4-(5-(4-chlorophenyl)-3-phenyl-1H-pyrazol-1-yl)benzenesulfonamide;

4-(3,5-bis(4-methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;

4-(5-(4-chlorophenyl)-3-(4-methylphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;

4-(5-(4-chlorophenyl)-3-(4-nitrophenyl)-1H-pyrazol-1-yl)benzenesulfonamide;

4-(5-(4-chlorophenyl)-3-(5-chloro-2-thienyl)-1H-pyrazol-1-yl)benzenesulfonamide;

4-(4-chloro-3,5-diphenyl-1H-pyrazol-1-yl)benzenesulfonamide

4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-phenyl-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-fluorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[4-chloro-5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-(4-methylphenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-phenyl-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-(4-methoxyphenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-cyano-5-(4-fluorophenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(3-fluoro-4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[4-chloro-5-phenyl-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-chlorophenyl)-3-(hydroxymethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-(N,N-dimethylamino)phenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

5-(4-fluorophenyl)-6-[4-(methylsulfonyl)phenyl]spiro[2.4]hept-5-ene;

4-[6-(4-fluorophenyl)spiro[2.4]hept-5-en-5-yl]benzenesulfonamide;

6-(4-fluorophenyl)-7-[4-(methylsulfonyl)phenyl]spiro[3.4]oct-6-ene;

5-(3-chloro-4-methoxyphenyl)-6-[4-(methylsulfonyl)phenyl]spiro[2.4]hept-5-ene;

4-[6-(3-chloro-4-methoxyphenyl)spiro[2.4]hept-5-en-5-yl]benzenesulfonamide;

5-(3,5-dichloro-4-methoxyphenyl)-6-[4-(methylsulfonyl)phenyl]spiro[2.4]hept-5-ene;

5-(3-chloro-4-fluorophenyl)-6-[4-(methylsulfonyl)phenyl]spiro[2.4]hept-5-ene;

4-[6-(3,4-dichlorophenyl)spiro[2.4]hept-5-en-5-yl]benzenesulfonamide;

2-(3-chloro-4-fluorophenyl)-4-(4-fluorophenyl)-5-(4-methylsulfonylphenyl)thiazole;

2-(2-chlorophenyl)-4-(4-fluorophenyl)-5-(4-methylsulfonylphenyl)thiazole;

5-(4-fluorophenyl)-4-(4-methylsulfonylphenyl)-2-methylthiazole;

4-(4-fluorophenyl)-5-(4-methylsulfonylphenyl)-2-trifluoromethylthiazole;

4-(4-fluorophenyl)-5-(4-methylsulfonylphenyl)-2-(2-thienyl)thiazole;

4-(4-fluorophenyl)-5-(4-methylsulfonylphenyl)-2-benzylaminothiazole;

4-(4-fluorophenyl)-5-(4-methylsulfonylphenyl)-2-(1-propylamino)thiazole;

2-[(3,5-dichlorophenoxy)methyl)-4-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]thiazole;

5-(4-fluorophenyl)-4-(4-methylsulfonylphenyl)-2-trifluoromethylthiazole;

1-methylsulfonyl-4-[1,1-dimethyl-4-(4-fluorophenyl)cyclopenta-2,4-dien-3-yl]benzene;

4-[4-(4-fluorophenyl)-1,1-dimethylcyclopenta-2,4-dien-3-yl]benzenesulfonamide;

5-(4-fluorophenyl)-6-[4-(methylsulfonyl)phenyl]spiro[2.4]hepta-4,6-diene;

4-[6-(4-fluorophenyl)spiro[2.4]hepta-4,6-dien-5-yl]benzenesulfonamide;

6-(4-fluorophenyl)-2-methoxy-5-[4-(methylsulfonyl)phenyl]-pyridine-3-carbonitrile;

2-bromo-6-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]-pyridine-3-carbonitrile;

6-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]-2-phenyl-pyridine-3-carbonitrile;

4-[2-(4-methylpyridin-2-yl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

4-[2-(5-methylpyridin-3-yl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

4-[2-(2-methylpyridin-3-yl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

3-[1-[4-(methylsulfonyl)phenyl]-4-(trifluoromethyl)-1H-imidazol-2-yl]pyridine;

2-[1-[4-(methylsulfonyl)phenyl-4-(trifluoromethyl)-1H-imidazol-2-yl]pyridine;

2-methyl-4-[1-[4-(methylsulfonyl)phenyl-4-(trifluoromethyl)-1H-imidazol-2-yl]pyridine;

2-methyl-6-[1-[4-(methylsulfonyl)phenyl-4-(trifluoromethyl)-1H-imidazol-2-yl]pyridine;

4-[2-(6-methylpyridin-3-yl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

2-(3,4-difluorophenyl)-1-[4-(methylsulfonyl)phenyl]-4-(trifluoromethyl)-1H-imidazole;

4-[2-(4-methylphenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

2-(4-chlorophenyl)-1-[4-(methylsulfonyl)phenyl]-4-methyl-1H-imidazole;

2-(4-chlorophenyl)-1-[4-(methylsulfonyl)phenyl]-4-phenyl-1H-imidazole;

2-(4-chlorophenyl)-4-(4-fluorophenyl)-1-[4-(methylsulfonyl)phenyl]-1H-imidazole;

2-(3-fluoro-4-methoxyphenyl)-1-[4-(methylsulfonyl)phenyl-4-(trifluoromethyl)-1H-imidazole;

1-[4-(methylsulfonyl)phenyl]-2-phenyl-4-trifluoromethyl-1H-imidazole;

2-(4-methylphenyl)-1-[4-(methylsulfonyl)phenyl]-4-trifluoromethyl-1H-imidazole;

4-[2-(3-chloro-4-methylphenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

2-(3-fluoro-5-methylphenyl)-1-[4-(methylsulfonyl)phenyl]-4-(trifluoromethyl)-1H-imidazole;

4-[2-(3-fluoro-5-methylphenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

2-(3-methylphenyl)-1-(4-(methylsulfonyl)phenyl]-4-trifluoromethyl-1H-imidazole;

4-[2-(3-methylphenyl)-4-trifluoromethyl-1H-imidazol-1-yl]benzenesulfonamide;

1-[4-(methylsulfonyl)phenyl]-2-(3-chlorophenyl)-4-trifluoromethyl-1H-imidazole;

4-[2-(3-chlorophenyl)-4-trifluoromethyl-1H-imidazol-1-yl]benzenesulfonamide;

4-[2-phenyl-4-trifluoromethyl-1H-imidazol-1-yl]benzenesulfonamide;

4-[2-(4-methoxy-3-chlorophenyl)-4-trifluoromethyl-1H-imidazol-1-yl]benzenesulfonamide;

1-allyl-4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-5-(trifluoromethyl)-1H-pyrazole;

4-[1-ethyl-4-(4-fluorophenyl)-5-(trifluoromethyl)-1H-pyrazol-3-yl]benzenesulfonamide;

N-phenyl-[4-(4-luorophenyl)-3-[4-(methylsulfonyl)phenyl]-5-(trifluoromethyl)-1H-pyrazol-1-yl]acetamide;

ethyl[4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-5-(trifluoromethyl)-1H-pyrazol-1-yl]acetate;

4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-1-(2-phenylethyl)-1H-pyrazole;

4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-1-(2-phenylethyl)-5-(trifluoromethyl)pyrazole;

1-ethyl-4-(4-fluorophenyl)-3-(4-(methylsulfonyl)phenyl]-5-(trifluoromethyl)-1H-pyrazole;

5-(4-fluorophenyl)-4-(4-methylsulfonylphenyl)-2-trifluoromethyl-1H-imidazole;

4-[4-(methylsulfonyl)phenyl]-5-(2-thiophenyl)-2-(trifluoromethyl)-1H-imidazole;

5-(4-fluorophenyl)-2-methoxy-4-[4-(methylsulfonyl)phenyl]-6-(trifluoromethyl)pyridine;

2-ethoxy-5-(4-fluorophenyl)-4-[4-(methylsulfonyl)phenyl]-6-(trifluoromethyl)pyridine;

5-(4-fluorophenyl)-4-[4-(methylsulfonyl)phenyl]-2-(2-propynyloxy)-6-(trifluoromethyl)pyridine;

2-bromo-5-(4-fluorophenyl)-4-[4-(methylsulfonyl)phenyl]-6-(trifluoromethyl)pyridine;

4-[2-(3-chloro-4-methoxyphenyl)-4,5-difluorophenyl]benzenesulfonamide;

1-(4-fluorophenyl)-2-[4-(methylsulfonyl)phenyl]benzene;

5-difluoromethyl-4-(4-methylsulfonylphenyl)-3-phenylisoxazole;

4-[3-ethyl-5-phenylisoxazol-4-yl]benzenesulfonamide;

4-[5-difluoromethyl-3-phenylisoxazol-4-yl]benzenesulfonamide;

4-[5-hydroxymethyl-3-phenylisoxazol-4-yl]benzenesulfonamide;

4-[5-methyl-3-phenyl-isoxazol-4-yl]benzenesulfonamide;

1-[2-(4-fluorophenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(4-fluoro-2-methylphenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(4-chlorophenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(2,4-dichlorophenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(4-trifluoromethylphenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(4-methylthiophenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(4-fluorophenyl)-4,4-dimethylcyclopenten-1-yl]-4-(methylsulfonyl)benzene;

4-[2-(4-fluorophenyl)-4,4-dimethylcyclopenten-1-yl]benzenesulfonamide;

1-[2-(4-chlorophenyl)-4,4-dimethylcyclopenten-1-yl]-4-(methylsulfonyl)benzene;

4-[2-(4-chlorophenyl)-4,4-dimethylcyclopenten-1-yl]benzenesulfonamide;

4-[2-(4-fluorophenyl)cyclopenten-1-yl]benzenesulfonamide;

4-[2-(4-chlorophenyl)cyclopenten-1-yl]benzenesulfonamide;

1-[2-(4-methoxyphenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

1-[2-(2,3-difluorophenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

4-[2-(3-fluoro-4-methoxyphenyl)cyclopenten-1-yl]benzenesulfonamide;

1-[2-(3-chloro-4-methoxyphenyl)cyclopenten-1-yl]-4-(methylsulfonyl)benzene;

4-[2-(3-chloro-4-fluorophenyl)cyclopenten-1-yl]benzenesulfonamide;

4-[2-(2-methylpyridin-5-yl)cyclopenten-1-yl]benzenesulfonamide;

ethyl2-[4-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]oxazol-2-yl]-2-benzyl-acetate;

2-[4-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]oxazol-2-yl]aceticacid;

2-(tert-butyl)-4-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]oxazole;

4-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]-2-phenyloxazole;

4-(4-fluorophenyl)-2-methyl-5-[4-(methylsulfonyl)phenyl]oxazole; and

4-[5-(3-fluoro-4-methoxyphenyl)-2-trifluoromethyl-4-oxazolyl]benzenesulfonamide.

A family of specific compounds of more particular interest withinFormula I consists of compounds and pharmaceutically-acceptable saltsthereof as follows:

4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(3-fluoro-4-methoxyphenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

3-[1-[4-(methylsulfonyl)phenyl]-4-trifluoromethyl-1H-imidazol-2-yl]pyridine;

2-methyl-5-[1-[4-(methylsulfonyl)phenyl]-4-trifluoromethyl-1H-imidazol-2-yl]pyridine;

4-[2-(5-methylpyridin-3-yl)-4-(trifluoromethyl)-1H-imidazol-1-yl]benzenesulfonamide;

4-[5-methyl-3-phenylisoxazol-4-yl]benzenesulfonamide;

4-[5-hydroxymethyl-3-phenylisoxazol-4-yl]benzenesulfonamide;

[2-trifluoromethyl-5-(3,4-difluorophenyl)-4-oxazolyl]benzenesulfonamide;

4-[2-methyl-4-phenyl-5-oxazolyl]benzenesulfonamide; and

4-[5-(3-fluoro-4-methoxyphenyl-2-trifluoromethyl)-4-oxazolyl]benzenesulfonamide.

A subclass of cyclooxygenase-2 inhibitors is selected from compounds ofWO95/15316. Preferably, the cyclooxygenase-2 inhibitor is selected fromcompounds of Formula II

wherein R⁴ is lower haloalkyl; wherein R⁵ is hydrido; and wherein R⁶ isphenyl optionally substituted at a substitutable position with one ormore radicals selected from halo, lower alkylthio, lower alkylsulfonyl,cyano, nitro, lower haloalkyl, lower alkyl, hydroxyl, lower alkenyl,lower hydroxyalkyl, carboxyl, lower cycloalkyl, lower alkylamino, lowerdialkylamino, lower alkoxycarbonyl, aminocarbonyl, lower alkoxy, lowerhaloalkoxy, sulfamyl, five or six membered heterocyclic and amino; or apharmaceutically-acceptable salt or derivative thereof.

A family of specific compounds of particular interest within Formula IIconsists of compounds, pharmaceutically-acceptable salts and derivativesthereof as follows:

4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-phenyl-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-fluorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-(4-methylphenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-phenyl-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-(4-methoxyphenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[3-(difluoromethyl)-5-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(3-fluoro-4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;and

4-[5-(4-(N,N-dimethylamino)phenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide.

A family of specific compounds of more particular interest withinFormula II consists of compounds and pharmaceutically-acceptable saltsor derivatives thereof as follows:

4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;

4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;and

4-[5-(3-fluoro-4-methoxyphenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide.

Derivatives are intended to encompass any compounds which arestructurally related to the cyclooxygenase-2 inhibitors or which possessthe substantially equivalent biologic activity. By way of example, suchinhibitors may include, but are not limited to, prodrugs thereof.

The compounds utilized in the methods of the present invention may bepresent in the form of free bases or pharmaceutically acceptable acidaddition salts thereof. The term “pharmaceutically-acceptable salts”embraces salts commonly used to form alkali metal salts and to formaddition salts of free acids or free bases. The nature of the salt isnot critical, provided that it is pharmaceutically-acceptable. Suitablepharmaceutically-acceptable acid addition salts of compounds of FormulaI may be prepared from an inorganic acid or from an organic acid.Examples of such inorganic acids are hydrochloric, hydrobromic,hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. Appropriateorganic acids may be selected from aliphatic, cycloaliphatic, aromatic,araliphatic, heterocyclic, carboxylic and sulfonic classes of organicacids, example of which are formic, acetic, propionic, succinic,glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic,anthranilic, mesylic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic(pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic,2-hydroxyethanesulfonic, toluenesulfonic, sulfanilic,cyclohexylaminosulfonic, stearic, algenic, β-hydroxybutyric, salicylic,galactaric and galacturonic acid. Suitable pharmaceutically-acceptablebase addition salts of compounds of Formula I include metallic saltsmade from aluminum, calcium, lithium, magnesium, potassium, sodium andzinc or organic salts made from chloroprocaine, choline,N,N′-dibenzylethylenediamine, diethanolamine, ethylenediamine, meglumine(N-methylglucamine) and procaine. All of these salts may be prepared byconventional means from the corresponding compound of Formula I byreacting, for example, the appropriate acid or base with the compound ofFormula I.

Biological Evaluation

The efficacy of cyclooxygenase-2 inhibitors as antineoplasia agents wasdetermined in the following models:

Murine Lewis lung Carcinoma Model.

Lewis lung carcinomas were implanted sub-cutaneously into the foot padof male C57BL/6 mice. The mice were subsequently treated with4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide.The drug was supplied in the drinking water at 6 mg/kg/day. Also anon-selective COX-1/COX-2 inhibitor indomethacin was tested in thismodel. The drug was supplied in the drinking water at the maximumtolerated dose of 2 mg/kg/day. A total of 10 mice/compound were tested.Tumor volume was determined twice a week using a plethysmometer. Theefficacy of these compounds on tumor growth was measured at day 32 aftercancer cell injection, as indicated in Table 1. The % inhibition valueis calculated by calculating the difference in tumor size compared withthe control group.

TABLE 1 Tumor Volume (Day 32) Treatment % Inhibition Vehicle/control0.00 COX-2 inhibitor 70.86 Indomethacin 62.90

Human prostate cancer cell tumors

Two human prostate cancer cell lines (PC-3 and LNCaP) were obtained(ATCC) to determine the efficacy of cyclooxygenase-2 inhibitors toinhibit tumor growth in a therapeutic model. In addition, the LNCaP cellline secretes prostate serum antigen (PSA) when grown in nude mice.

PC-3

PC-3 cells (10⁶ cells/0.2 ml of 30% matrigel) in RPMI 1640 medium wasinjected on the back of nude mice. At day 28, a COX-2 inhibitor4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide(20 mg/kg/day in water) was administered. After 45 days, PGE₂ and TXB₂were measured. The COX-2 inhibitor inhibited tumor growth by 55%. PGE₂and TXB₂ levels were reduced by 80-90% in the animals treated with theCOX-2 inhibitor.

LNCaP

Similar to the results in PC-3, a COX-2 inhibitor4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamideat a dose equivalent to 6 mg/kg/day in the drinking water inhibited thegrowth of the tumor by 55% at day 58. PSA level was reduced toapproximately 50% as judged by western blotting.

Others

Cell lines: The following cell lines can be used: classic small celllung cancer (SCLC) cell lines NCI-H209, NCI-H345, and NCI-H510; variantSCLC cell lines NCI-N417 and NCI-H82; large cell carcinoma cell lineNCI-H1155; adeno carcinoma cell line NCI-H23; and bronchioalveolearcarcinoma cell line A549, breast cancer cell line MCF-7 (American TypeTissue Culture Rockville MD; ATCC) and colon cancer cell lines such asNCI-H630 (ATCC), HT 29, SW948, HCA-7 and others that can be tested invivo or in vitro. All cells can be grown in RPMI-1640, supplemented with5% fetal bovine serum (FBS), penicillin and streptomycin (Gibco, GrandIsland, N.Y.), and be maintained in a 5% CO₂ atmosphere at 37° C. Allcell lines are free of mycoplasma contamination.

Growth studies: A modification (Promega CellTiter 96®, Promega Madison,Wis.) of the semiautomated colorimetric assay, MTT [Nakanishi, et al.Exper. Cell Biol., 56, 74-85 (1988)], which quantitates cell numbersbased on reduction of a tetrazolium compound by tumor cells asdetermined by a spectrophotometer (540 nm) is used. All assays areperformed in RPMI-1640 media supplemented with transfertin ˜10 g/ml,insulin ˜5 g/ml and selenium (Sigma Chemicals, St. Louis, Mo.). Seedingdensities are ˜2×10⁴ cells/well, and cells are grown for 5 days. Eachexperiment is reported as mean optical density corrected for background+/− standard deviation. The cyclooxygenasse-2 inhibitors should beactive, at a dose of 20 mg/kg, in inhibiting growth of the cancerouscell lines.

A mouse urinary bladder tumor model is performed with materials,reagents and procedures essentially as described by Grubbs et al,[Anticancer Res., 13, 33-36 (1993)]. A COX-2 inhibitor should be activeat a dose of 20 mg/kg.

A rat mammary tumor model is performed with materials, reagents andprocedures essentially as described by Grubbs et al., [Anticancer Res.,15, 709-16 (1995)]. A COX-2 inhibitor should be active at a dose of 20mg/kg.

A mouse cervical and vaginal carcinogenesis model is performed withmaterials, reagents and procedures essentially as described by Arbeit etal., [Proc. Acad. Sci. USA., 93, 2930-35 (1996)]. A COX-2 inhibitorshould be active at a dose of 20 mg/kg.

A colon adenocarcinoma cell model is performed with materials, reagentsand procedures essentially as described by Shiff et al., [J. Clin.Invest., 96, 491-503 (1995)]. A COX-2 inhibitor should be active at adose of 20 mg/kg. See also Masahiko Tsujii et al. (Proc. Natl. Acad.Sci. USA 94:3336-3340, 1997).

In summary, COX-2 inhibitors reduce tumor growth in several animalcancer models.

Combination Therapy of a COX-2 inhibitor and other antineoplastic agents

Lewis Lung carcinoma cells (2.5×10⁶ cells) prepared from a brei carriedin C57BL/6 mice were injected subcutaneously into the hind legs of mice.A COX-2 inhibitor,4-[5-(4-chlorophenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamidewas given by gavage twice a week to groups of 10 mice at doses of 6 and20 mg/kg. Cyclophosphamide (CTX) was injected to mice on days 5,7 and 9after the implantation of the tumor at a dose of 50 mg/kg. Tumor volumewas determined during the study. Animals were sacrificed at day 26 andthe results of this experiments are summarized in Table 2. The %inhibition was calculated as above.

TABLE 2 Tumor Volume (Day 22) Treatment % Inhibition Vehicle 0 COX-2inhibitor (6 mg/kg) 0 COX-2 inhibitor (20 mg/kg) 54 CTX (50 mg/kg) 57CTX + COX-2 inhibitor (6 mg/kg) 69 CTX + COX-2 inhibitor (20 mg/kg) 77

The results of this experiment indicate that the combination of a COX-2inhibitor and a cytotoxic agent produced an additive effect on theirindividual capacity to inhibit tumor growth.

The active compounds of the present invention may be administered by anysuitable route known to those skilled in the art, preferably in the formof a pharmaceutical composition adapted to such a route, and in a doseeffective for the treatment intended. The active compounds andcomposition may, for example, be administered orally, intravascularly,intraperitoneally, intranasal, intrabronchial, subcutaneously,intramuscularly or topically (including aerosol).

The administration of the present invention may be for either preventionor treatment purposes. The methods and compositions used herein may beused alone or in conjunction with additional therapies known to thoseskilled in the art in the prevention or treatment of neoplasia.Alternatively, the methods and compositions described herein may be usedas conjunctive therapy. By way of example, the cyclooxygenase-2inhibitor may be administered alone or in conjunction with otherantineoplastic agents or other growth inhibiting agents or other drugsor nutrients.

There are large numbers of antineoplastic agents available in commercialuse, in clinical evaluation and in pre-clinical development, which couldbe selected for treatment of neoplasia by combination drug chemotherapy.Such antineoplastic agents fall into several major categories, namely,antibiotic-type agents, alkylating agents, antimetabolite agents,hormonal agents, immunological agents, interferon-type agents and acategory of miscellaneous agents. Alternatively, other anti-neoplasticagents, such as metallomatrix proteases (MMP), SOD mimics or α_(v)β₃inhibitors may be used.

A first family of antineoplastic agents which may be used in combinationwith a selective cyclooxygenase-2 inhibitor consists ofantimetabolite-type antineoplastic agents. Suitable antimetaboliteantineoplastic agents may be selected from the group consisting of5-FU-fibrinogen, acanthifolic acid, aminothiadiazole, brequinar sodium,carmofur, Ciba-Geigy CGP-30694, cyclopentyl cytosine, cytarabinephosphate stearate, cytarabine conjugates, Lilly DATHF, Merrel Dow DDFC,dezaguanine, dideoxycytidine, dideoxyguanosine, didox, Yoshitomi DMDC,doxifluridine, Wellcome EHNA, Merck & Co. EX-015, fazarabine,floxuridine, fludarabine phosphate, 5-fluorouracil,N-(2′-furanidyl)-5-fluorouracil, Daiichi Seiyaku FO-152, isopropylpyrrolizine, Lilly LY-188011, Lilly LY-264618, methobenzaprim,methotrexate, Wellcome MZPES, norspermidine, NCI NSC-127716, NCINSC-264880, NCI NSC-39661, NCI NSC-612567, Warner-Lambert PALA,pentostatin, piritrexim, plicamycin, Asahi Chemical PL-AC, TakedaTAC-788, thioguanine, tiazofurin, Erbamont TIF, trimetrexate, tyrosinekinase inhibitors, tyrosine protein kinase inhibitors, Taiho UFT anduricytin.

A second family of antineoplastic agents which may be used incombination with a selective cyclooxygenase-2 inhibitor consists ofalkylating-type antineoplastic agents. Suitable alkylating-typeantineoplastic agents may be selected from the group consisting ofShionogi 254-S, aldo-phosphamide analogues, altretamine, anaxirone,Boehringer Mannheim BBR-2207, bestrabucil, budotitane, Wakunaga CA-102,carboplatin, carmustine, Chinoin-139, Chinoin-153, chlorambucil,cisplatin, cyclophosphamide, American Cyanamid CL-286558, Sanofi CY-233,cyplatate, Degussa D-19-384, Sumimoto DACHP(Myr)2, diphenylspiromustine,diplatinum cytostatic, Erba distamycin derivatives, Chugai DWA-2114R,ITI E09, elmustine, Erbamont FCE-24517, estramustine phosphate sodium,fotemustine, Unimed G-6-M, Chinoin GYKI-17230, hepsul-fam, ifosfamide,iproplatin, lomustine, mafosfamide, mitolactol, Nippon Kayaku NK-121,NCI NSC-264395, NCI NSC-342215, oxaliplatin, Upjohn PCNU, prednimustine,Proter PTT-119, ranimustine, semustine, SmithKline SK&F-101772, YakultHonsha SN-22, spiromustine, Tanabe Seiyaku TA-077, tauromustine,temozolomide, teroxirone, tetraplatin and trimelamol.

A third family of antineoplastic agents which may be used in combinationwith a selective cyclooxygenase-2 inhibitor consists of antibiotic-typeantineoplastic agents. Suitable antibiotic-type antineoplastic agentsmay be selected from the group consisting of Taiho 4181-A, aclarubicin,actinomycin D, actinoplanone, Erbamont ADR-456, aeroplysinin derivative,Ajinomoto AN-201-II, Ajinomoto AN-3, Nippon Soda anisomycins,anthracycline, azino-mycin-A, bisucaberin, Bristol-Myers BL-6859,Bristol-Myers BMY-25067, Bristol-Myers BMY-25551, Bristol-MyersBMY-26605, Bristol-Myers BMY-27557, Bristol-Myers BMY-28438, bleomycinsulfate, bryostatin-1, Taiho C-1027, calichemycin, chromoximycin,dactinomycin, daunorubicin, Kyowa Hakko DC-102, Kyowa Hakko DC-79, KyowaHakko DC-88A, Kyowa Hakko DC89-A1, Kyowa Hakko DC92-B, ditrisarubicin B,Shionogi DOB-41, doxorubicin, doxorubicin-fibrinogen, elsamicin-A,epirubicin, erbstatin, esorubicin, esperamicin-A1, esperamicin-Alb,Erbamont FCE-21954, Fujisawa FK-973, fostriecin, Fujisawa FR-900482,glidobactin, gregatin-A, grincamycin, herbimycin, idarubicin, illudins,kazusamycin, kesarirhodins, Kyowa Hakko KM-5539, Kirin Brewery KRN-8602,Kyowa Hakko KT-5432, Kyowa Hakko KT-5594, Kyowa Hakko KT-6149, AmericanCyanamid LL-D49194, Meiji Seika ME 2303, menogaril, mitomycin,mitoxantrone, SmithKline M-TAG, neoenactin, Nippon Kayaku NK-313, NipponKayaku NKT-01, SRI International NSC-357704, oxalysine, oxaunomycin,peplomycin, pilatin, pirarubicin, porothramycin, pyrindamycin A, TobishiRA-I, rapamycin, rhizoxin, rodorubicin, sibanomicin, siwenmycin,Sumitomo SM-5887, Snow Brand SN-706, Snow Brand SN-07, sorangicin-A,sparsomycin, SS Pharmaceutical SS-21020, SS Pharmaceutical SS-7313B, SSPharmaceutical SS-9816B, steffimycin B, Taiho 4181-2, talisomycin,Takeda TAN-868A, terpentecin, thrazine, tricrozarin A, Upjohn U-73975,Kyowa Hakko UCN-10028A, Fujisawa WF-3405, Yoshitomi Y-25024 andzorubicin.

A fourth family of antineoplastic agents which may be used incombination with the selective cyclooxygenase-2 inhibitor consists of amiscellaneous family of antineoplastic agents selected from the groupconsisting of alpha-carotene, alpha-difluoromethyl-arginine, acitretin,Biotec AD-5, Kyorin AHC-52, alstonine, amonafide, amphethinile,amsacrine, Angiostat, ankinomycin, anti-neoplaston A10, antineoplastonA2, antineoplaston A3, antineoplaston A5, antineoplaston AS2-1, HenkelAPD, aphidicolin glycinate, asparaginase, Avarol, baccharin, batracylin,benfluron, benzotript, Ipsen-Beaufour BIM-23015, bisantrene,Bristo-Myers BMY-40481, Vestar boron-10, bromofosfamide, WellcomeBW-502, Wellcome BW-773, caracemide, carmethizole hydrochloride,Ajinomoto CDAF, chlorsulfaquinoxalone, Chemes CHX-2053, Chemex CHX-100,Warner-Lambert CI-921, Warner-Lambert CI-937, Warner-Lambert CI-941,Warner-Lambert CI-958, clanfenur, claviridenone, ICN compound 1259, ICNcompound 4711, Contracan, Yakult Honsha CPT-11, crisnatol, curaderm,cytochalasin B, cytarabine, cytocytin, Merz D-609, DABIS maleate,dacarbazine, datelliptinium, didemnin-B, dihaematoporphyrin ether,dihydrolenperone, dinaline, distamycin, Toyo Pharmar DM-341, ToyoPharmar DM-75, Daiichi Seiyaku DN-9693, elliprabin, elliptinium acetate,Tsumura EPMTC, ergotamine, etoposide, etretinate, fenretinide, FujisawaFR-57704, gallium nitrate, genkwadaphnin, Chugai GLA-43, Glaxo GR-63178,grifolan NMF-5N, hexadecylphosphocholine, Green Cross HO-221,homoharringtonine, hydroxyurea, BTG ICRF-187, ilmofosine, isoglutamine,isotretinoin, Otsuka JI-36, Ramot K-477, Otsuak K-76COONa, KurehaChemical K-AM, MECT Corp KI-8110, American Cyanamid L-623, leukoregulin,lonidamine, Lundbeck LU-23-112, Lilly LY-186641, NCI (US) MAP, marycin,Merrel Dow MDL-27048, Medco MEDR-340, merbarone, merocyaninederivatives, methylanilinoacridine, Molecular Genetics MGI-136,minactivin, mitonafide, mitoquidone, mopidamol, motretinide, ZenyakuKogyo MST-16, N-(retinoyl)amino acids, Nisshin Flour Milling N-021,N-acylated-dehydroalanines, nafazatrom, Taisho NCU-190, nocodazolederivative, Normosang, NCI NSC-145813, NCI NSC-361456, NCI NSC-604782,NCI NSC-95580, octreotide, Ono ONO-112, oquizanocine, Akzo Org-10172,pancratistatin, pazelliptine, Warner-Lambert PD-111707, Warner-LambertPD-115934, Warner-Lambert PD-131141, Pierre Fabre PE-1001, ICRT peptideD, piroxantrone, polyhaematoporphyrin, polypreic acid, Efamol porphyrin,probimane, procarbazine, proglumide, Invitron protease nexin I, TobishiRA-700, razoxane, Sapporo Breweries RBS, restrictin-P, retelliptine,retinoic acid, Rhone-Poulenc RP-49532, Rhone-Poulenc RP-56976,SmithKline SK&F-104864, Sumitomo SM-108, Kuraray SMANCS, SeaPharmSP-10094, spatol, spirocyclopropane derivatives, spirogermanium, Unimed,SS Pharmaceutical SS-554, strypoldinone, Stypoldione, Suntory SUN 0237,Suntory SUN 2071, superoxide dismutase, Toyama T-506, Toyama T-680,taxol, Teijin TEI-0303, teniposide, thaliblastine, Eastman Kodak TJB-29,tocotrienol, Topostin, Teijin TT-82, Kyowa Hakko UCN-01, Kyowa HakkoUCN-1028, ukrain, Eastman Kodak USB-006, vinblastine sulfate,vincristine, vindesine, vinestramide, vinorelbine, vintriptol,vinzolidine, withanolides and Yamanouchi YM-534.

Examples of radioprotective agents which may be used in the combinationchemotherapy of this invention are AD-5, adchnon, amifostine analogues,detox, dimesna, 1-102, MM-159, N-acylated-dehydroalanines,TGF-Genentech, tiprotimod, amifostine, WR-151327, FUT-187, ketoprofentransdermal, nabumetone, superoxide dismutase (Chiron) and superoxidedismutase Enzon.

Methods for preparation of the antineoplastic agents described above maybe found in the literature. Methods for preparation of doxorubicin, forexample, are described in U.S. Pat. No. 3,590,028 and No. 4,012,448.Methods for preparing metallomatrix protease inhibitors are described inEP 780386. Methods for preparing SOD mimics are described in EP 524,101.Methods for preparing α_(v)β₃ inhibitors are described in WO97/08174.

The phrase “conjunctive therapy” (or “combination therapy”), in defininguse of a cyclooxygenase-2 inhibitor agent and another pharmaceuticalagent, is intended to embrace administration of each agent in asequential manner in a regimen that will provide beneficial effects ofthe drug combination, and is intended as well to embraceco-administration of these agents in a substantially simultaneousmanner, such as in a single formulation having a fixed ratio of theseactive agents, or in multiple, separate formulations for each agent. Thepresent invention also comprises a pharmaceutical composition for theprevention and treatment of neoplasia, comprising atherapeutically-effective amount of a compound of Formula I inassociation with at least one pharmaceutically-acceptable carrier,adjuvant or diluent (collectively referred to herein as “carrier”materials) and, other antineoplastic agents or other growth inhibitingagents or other drugs or nutrients.

For oral administration, the pharmaceutical composition may be in theform of, for example, a tablet, capsule, suspension or liquid. Thepharmaceutical composition is preferably made in the form of a dosageunit containing a particular amount of the active ingredient. Examplesof such dosage units are capsules, tablets, powders, granules or asuspension, with conventional additives such as lactose, mannitol, cornstarch or potato starch; with binders such as crystalline cellulose,cellulose derivatives, acacia, corn starch or gelatins; withdisintegrators such as corn starch, potato starch or sodiumcarboxymethyl-cellulose; and with lubricants such as talc or magnesiumstearate. The active ingredient may also be administered by injection asa composition wherein, for example, saline, dextrose or water may beused as a suitable carrier.

For intravenous, intramuscular, subcutaneous, or intraperitonealadministration, the compound may be combined with a sterile aqueoussolution which is preferably isotonic with the blood of the recipient.Such formulations may be prepared by dissolving solid active ingredientin water containing physiologically compatible substances such as sodiumchloride, glycine, and the like, and having a buffered pH compatiblewith physiological conditions to produce an aqueous solution, andrendering said solution sterile. The formulations may be present in unitor multi-dose containers such as sealed ampoules or vials.

If the neoplasia is localized in the G.I. tract, the compound may beformulated with acid-stable, base-labile coatings known in the art whichbegin to dissolve in the high pH small intestine. Formulation to enhancelocal pharmacologic effects and reduce systemic uptake are preferred.

Formulations suitable for parenteral administration convenientlycomprise a sterile aqueous preparation of the active compound which ispreferably made isotonic. Preparations for injections may also beformulated by suspending or emulsifying the compounds in non-aqueoussolvent, such as vegetable oil, synthetic aliphatic acid glycerides,esters of higher aliphatic acids or propylene glycol.

Formulations for topical use include known gels, creams, oils, and thelike. For aerosol delivery, the compounds may be formulated with knownaerosol exipients, such as saline, and administered using commerciallyavailable nebulizers. Formulation in a fatty acid source may be used toenhance biocompatibility. Aerosol delivery is the preferred method ofdelivery for epithelial neoplasias of the lung for preventionapplication.

For rectal administration, the active ingredient may be formulated intosuppositories using bases which are solid at room temperature and meltor dissolve at body temperature. Commonly used bases include cocoabutter, glycerinated gelatin, hydrogenated vegetable oil, polyethyleneglycols of various molecular weights, and fatty esters of polyethylenestearate.

The dosage form and amount can be readily established by reference toknown neoplasia treatment or prophylactic regiments. The amount oftherapeutically active compound that is administered and the dosageregimen for treating a disease condition with the compounds and/orcompositions of this invention depends on a variety of factors,including the age, weight, sex and medical condition of the subject, theseverity of the disease, the route and frequency of administration, andthe particular compound employed, the location of the neoplasia, as wellas the pharmacokinetic properties of the individual treated, and thusmay vary widely. The dosage will generally be lower if the compounds areadministered locally rather than systemically, and for prevention ratherthan for treatment. Such treatments may be administered as often asnecessary and for the period of time judged necessary by the treatingphysician. One of skill in the art will appreciate that the dosageregime or therapeutically effective amount of the inhibitor to beadministrated may need to be optimized for each individual. Thepharmaceutical compositions may contain active ingredient in the rangeof about 0.1 to 2000 mg, preferably in the range of about 0.5 to 500 mgand most preferably between about 1 and 200 mg. A daily dose of about0.01 to 100 mg/kg body weight, preferably between about 0.1 and about 50mg/kg body weight, may be appropriate. The daily dose can beadministered in one to four doses per day.

All patents and documents referenced herein are incorporated byreference.

Although this invention has been described with respect to specificembodiments, the details of these embodiments are not to be construed aslimitations.

What is claimed is:
 1. A method of treating a neoplasia in a subject inneed of such treatment, said method comprising treating the subject witha therapeutically-effective amount of a compound of Formula II

or a pharmaceutically acceptable salt thereof wherein R¹ is methyl oramino; R⁴ is (i) C₁-C₆ haloalkyl, (ii) phenyl optionally substitutedwith one or more members independently selected from the groupconsisting of C₁-C₆ alkyl, C₁-C₆ alkoxy, nitro, cyano, halogen, hydroxy,amino, C₁-C₆ alkyl amino, and di-C₁-C₆ alkylamino groups, or (iii)thienyl optionally substituted with one or more members independentlyselected from the group consisting of C₁-C₆ alkyl, C₁-C₆ alkoxy, nitro,cyano, halogen, hydroxy, amino, C₁-C₆ alkylamino, anddi(C₁-C₆)alkylamino groups; and R⁶ is phenyl optionally substituted at asubstitutable position with one or more radicals selected from the groupconsisting of halo, C₁-C₆ alkylthio, C₁-C₆ alkylsulfonyl, cyano, nitro,C₁-C₆ haloalkyl, C₁-C₆ alkyl, hydroxyl, C₁-C₆ alkenyl, C₁-C₆hydroxyalkyl, carboxyl, C₁-C₆ cycloalkyl, C₁-C₆ alkylamino, C₁-C₆dialkylamino, C₁-C₆ alkoxycarbonyl, aminocarbonyl, C₁-C₆ alkoxy, C₁-C₆haloalkoxy, sulfamyl, five or six membered heterocyclic radicals, andamino.
 2. The method of claim 1 wherein the neoplasia is selected fromthe group consisting of colorectal cancer, gastrointestinal cancer,liver cancer, bladder cancer, cervical cancer, prostate cancer, lungcancer, breast cancer and skin cancer.
 3. A method of claim 1, whereinthe neoplasia is adenomatous polyps.
 4. A method of treating a subjectsuffering from a neoplastic disease state with a conjunctive therapy,said method comprising treating the subject with atherapeutically-effective amount of a cyclooxygenase-2 selectivecompound and a compound selected from the group consisting of antibioticagents, alkylating agents, antimetabolite agents, hormonal agents,immunological agents, interferon agents, metallomatrix proteases (MMP)inhibitors, SOD, and α_(v)β₃ inhibitors, wherein the selective COX-2inhibitor is a compound of formula II or formula III

or a pharmaceutically acceptable salt thereof wherein each R¹ is methylor amino; R⁴ is (i) C₁-C₆ haloalkyl, (ii) phenyl optionally substitutedwith one or more members independently selected from the groupconsisting of C₁-C₆ alkyl, C₁-C₆ alkoxy, nitro, cyano, halogen, hydroxy,amino, C₁-C₆ alkyl amino, and di-C₁-C₆ alkylamino groups, or (iii)thienyl optionally substituted with one or more members independentlyselected from the group consisting of C₁-C₆ alkyl, C₁-C₆ alkoxy, nitro,cyano, halogen, hydroxy, amino C₁-C₆ alkylamino, and di(C₁-C₆)alkylamino groups; each R⁶ is phenyl optionally substituted at asubstitutable position with one or more radicals selected from the groupconsisting of halo, C₁-C₆ alkythio, C₁-C₆ alkylsulfonyl, cyano, nitro,C₁-C₆ haloalkyl, C₁-C₆ alkyl, hydroxyl, C₁-C₆ alkenyl, C₁-C₆HYDROXYALKYL, carboxyl, C₁-C₆ cycloalkyl, C₁-C₆ alkylamino, C₁-C₆dialkylamino, C₁-C₆ alkoxycarbonyl, aminocarbonyl, C₁-C₆ alkoxy, C₁-C₆haloalkoxy, sulfamyl, five or six membered heterocyclic radicals, andamino; R⁸ hydrogen or C₁-C₆ haloalkyl; and R⁹ is C₁-C₆ alkenyl, C₁-C₆alkoxycarbonyl(C₁-C₂) alkyl N-phenylacetamido, or phenyl(C₁-C₆)alkyl. 5.The method of claim 4 wherein the compound is selected from compounds,and their pharmaceutically acceptable salts, of the group consisting of5-(4-flurophenyl)-1-[4-(methysulfonyl)phenyl]-3-(trifluromethyl)pyrazole;4-(4-flurophenyl)-5-[4-(methysulfonyl)phenyl]-1-phenyl-3-(trifluoromethyl)pyrazole;4-(5-(4-chlorophenyl)-3-(4-methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(3,5-bis(4-methyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(5-(4-chlorophenyl)-3-phenyl-1H-pyrazol-1-yl)benzenesulfonamide;4-(3,5-bis(4-methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(5-(4-chlorophenyl)-3-(4-methlphenyl)-1H-pyrazol-1-yl)benezenesulfonamide;4-(5-(4-chlorophenyl)-3-(4-nitrophenyl)-1H-pyrazol-1-yl)benezenesulfonamide;4-(5-(4-chlorophenyl)-3-(5-chloro-2-thienyl)-1H-pyrazol-1-yl)benzensulfonamide;4-(4-chloro-3,5-diphenyl-1H-pyrazol-1-yl)benzenesulfonamide;4-[4-chloro-5-(4-chlorophenyl)-3-(trifluoromethyl]-1H-pyrazol-1yl)benezensulfonamide;4-[3-cyano-5-(4-fluorophenyl)-1H-pyrazol-1-yl]benzenesulfonamide;4-[4-chloro-5-phenyl-1H-pyrazol-1-yl]benzenesulfonamide;4-[(5-(4-chlorophenyl)-3-(hydroxymethyl)-1H-pyrazol-1-yl]benzenesulfonamide;1-allyl-4-(4-flurophenyl)-3-(4-(methysulsonyl)phenyl)-5-(trifluoromethyl)-1H-pyrazole;4-[1-ethyl-4-(4-fluorophenyl)-5-(trifluoromethyl)1H-pyrazol-3yl]benzenesulfonamide;N-phenyl-[4-(4-fluorophenyl)-3-(4-(methyysulfonyl)phenyl)-5-(trifluromethyl)-1H-pyrazol-1yl]acetamide;ethyl[4-(4-fluorophenyl)-3-(4-methysulfonyl)phenyl)-5-(trifluoromethyl)-1H-pyrazol-1-yl]acetate;4-(4-fluorophenyl)-3-[4-methysulfonyl)phenyl]-1-(2-phenylethyl)-1H-pyrazole;4-(4-fluorophenyl)-3-[4-methysulfonyl]phenyl)-1-(2-phenylethyl)-5-(trifluoromethyl)pyrazole;and1-ethyl-4-(4-fluorophenyl)-3-(4-(methysulfonyl)phenyl)-5-(trifluoromethyl)-1H-pyrazole.6. The method of claim 5 wherein the compound is selected fromcompounds, and their pharmaceutically acceptable salts, of the groupconsisting of4-[5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;4-[5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;and4-[5-(3-fluoro-4-methoxyphenyl)-3-(difluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide.7. A method of treating a neoplasia in a subject in need of suchtreatment, said method comprising treating the subject with atherapeutically-effective amount of a compound of the formula

or a pharmaceutically acceptable salt thereof wherein R¹ is methyl oramino; R⁶ is phenyl optionally substituted at a substitutable positionwith one or more radicals selected from the group consisting of halo,C₁-C₆ alkylthio, C₁-C₆ alkylsulfonyl, cyano, nitro, C₁-C₆ haloalkyl,C₁-C₆ alkyl, hydroxyl, C₁-C₆ alkenyl, C₁-C₆ hydroxyalkyl, carboxyl,C₁-C₆ cyloalalkyl, C₁-C₆ alkylamico, C₁-C₆ dialkylamino, C₁-C₆alkoxycarbonyl, aminocarbonyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, sulfamyl,five or six membered heterocyclic radicals, and amino; R⁸ is hydrogen orC₁-C₆ hyloalkyl; and R⁹ C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxycarbonyl(C₁-C₂) alkyl, N-phenylacetamido, or phenyl (C₁-C₆) alkyl.
 8. The methodof claim 7 wherein the compound is selected from compounds, and theirpharmaceutically acceptable salts, of the group consisting of5-(4-fluorophenyl)-1-4-(methylsulfonyl)phenyl)-3-(trifluoromethyl)pyrazole;4-(4-fluorophenyl)-5-4-(methylsulfonyl)phenyl)-1-phenyl-3-(trifluoromethyl)pyrazole;4-(5-(4-chlorophenyl)-3-(4-methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(3,5-bis(4-methylphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(5-(4-chlorophenyl)-3-phenyl-1H-pyrazol-1-yl)benzenesulfonamide;4-(3,5-bis(4-methoxyphenyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(5-(4-chlorophenyl)-3-(4-methylphenyl)-1H-pyrazol-1-pyrazol)benzenesulfonamide;4-(5-(4-chlorophenyl)-3-(4-nitrophenyl)-1H-pyrazol-1yl)benzenesulfonamide;4-(5-(4-chlorophenyl)-3-(5-chloro-2-thienyl)-1H-pyrazol-1-yl)benzenesulfonamide;4-(4-chloro-3,5-diphenyl-1H-pyrazol-1-yl)benzenesulfonamide;4-[4-chloro-5-(4-chlorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]benzenesulfonamide;4-[3-cyano-5-(4-fluorophenyl)-1H-pyrazol-1-yl]benzenesulfonamide;4-[4-chloro-5-phenyl-1H-pyrazol-1-yl]benzenesulfonamide; and4-[(5-(4-chlorophenyl)-3-(hydroxymethyl)-1H-pyrazol-1-yl]benzenesulfonamide.9. The method of claim 7 wherein the compound is selected fromcompounds, and their pharmaceutically acceptable salts, of the groupconsisting of1-allyl-4-(4-fluorophenyl)-3-[4-(methysulfonyl)phenyl]-5-(trifluoromethyl)-1H-pyrazole;4-[1-ethyl-4-(4-fluorophenyl)-5-(trifluoromethyl)-1H-pyrazol-3-yl]benzenesulfonamide;N-phenyl-4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl-5-(trifluromethyl)-1H-pyrazol-1-yl]acetamide;ethyl4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl-5-(trifluoromethyl)-1H-pyrazol-1-yl]acetate;4-(4-fluorophenyl)-3-4-[(methylsulfonyl)phenyl]-1-(2-phenylethyl)-1H-pyrazole;4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-1-(2-phenylethyl)-5-(trifluoromethyl)pyrazole;and1-ethyl-4-(4-fluorophenyl)-3-[4-(methylsulfonyl)phenyl]-5-(trifluoromethyl)-1H-pyrazole.10. A method of treating familial adenomatous polyps in a subject inneed of such treatment, said method comprising administering to thesubject a therepeutically-effective amount of a compound which is4-5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzenesulfonamide.11. A method of treating familial adenomatous polyps in a subject inneed of such treatment, said method comprising admistering to thesubject a pharmaceutical composition comprising4-5-(4-methylphenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzenesulfonamidetogether with a pharmaceutically acceptable carrier.
 12. A methodaccording to claim 11, wherein the carrier comprises lactose, magnesiumstearate, and gelatin.